Evaluation of usefulness of Microbial Assay for Risk Assessment (MARA) in the cyanobacterial toxicity estimation

The aim of the study was to elucidate the usefulness of the Microbial Assay for Risk Assessment (MARA) to evaluate toxicity in samples containing cyanobacterial products. Cyanobacterial extracts with different cyanotoxin contents and pure cyanotoxins—microcystin-LR, cylindrospermopsin and anatoxin-a—were tested. On the basis of the microbial reaction, MARA indicated only slight or no toxicity in the studied extracts. Similarly, no or low toxicity of pure toxins was detected at the concentrations used (up to 10 μg/ml). Weak relationships between the reactions of individual organisms exposed to cyanotoxin-containing extracts and to the same pure toxins were observed. On the other hand, inhibition of some organisms, such as Pichia anomalia, whose growth was not impacted by pure cyanotoxins, indicated the presence of other biologically active compounds in the studied extracts. In conclusion, MARA assay is not enough sensitive to be used as a good tool for cyanotoxin screening. It may, however, be applied in searching for antimicrobial/antifungal cyanobacteria-derived compounds.     

Results of a technology demonstration project to compare rapid aquatic toxicity screening tests in the analysis of industrial effluents

The results of a 'BioWise' demonstration project to assess the comparative sensitivity and practicality of seven new assays for the direct assessment of ecotoxicity in industrial effluents are presented. In addition the aim of the project was to validate the results of the new assays against benchmark data generated from non-proprietary, rapid, microplate screening assays using the regulatory species; freshwater crustacean Daphnia magna and green algae Selenastrum capricornutum, chosen in view of their environmental relevance. The new commercial test assays were: Daphnia magna, Selenastrum capricornutum and Thamnocephalus platyurus Toxkits supplied by Vickers Laboratories Ltd, containing dormant, immobilised life stages of the test species; GreenScreen EM, a yeast based assay for genotoxicity and general acute toxicity supplied by Gentronix Ltd; and CellSense a mediated, amperometric whole cell biosensor based on immobilised activated sludge and E. coli. 38 effluent samples supplied by members of SOCSA (Specialised Organic Chemicals Sector Association) were examined over a period of 13 months, in the project co-ordinated by the AstraZeneca Brixham Environmental Laboratory, and part funded by BioWise via the UK Government Department of Trade and Industry.     

A simple,cost-effective multispecies toxicity test using organisms with a cosmopolitan distribution

Difficulties in making accurate, ecosystem-level predictions of environmental effects of chemicals, mixtures, and effluents based solely on the results of tests on single species have necessitated the development of more environmentally realistic, predictive testing methods. This paper describes a multispecies, community-level toxicity test based on the colonization of artificial substrates by microbial species. Tests examined the colonization of initially barren polyurethane foam artificial substrates by Protozoa from a species source colonized in a natural system. Differences in colonization were examined in microecosystems amended with low levels of cadmium, a very toxic heavy metal, and TFM, an organic biocide used against larval sea lamprey. Tests examined differences in colonization over 28 days. For cadmium, effect levels were estimated to be near 1 g 1^–1, in the low range of effect levels determined from chronic single species tests. For TFM, effect levels were estimated to be between 1 and 10 ppm, overlapping the concentrations used in environmental applications. The colonization response, which depends on naked microbial cells reproducing and migrating through toxicant amended water to new substrates, is very sensitive. Tests based on colonization can be adapted to use species from a target receiving system or can use species from a designated natural source. Field validation of these tests can employ nearly identical methods to those used in laboratory studies to assess the accuracy of predictions based on test system data.     

Preliminary results of laboratory toxicity tests with the mayfly, Isonychia bicolor (Ephemeroptera: Isonychiidae) for development as a standard test organism for evaluating streams in the Appalachian coalfields of Virginia and West Virginia

Selecting the most appropriate test species for sediment and water column assays has been a primary goal for ecotoxicologists. Standard test organisms and established test guidelines exist, but the USEPA-recommended species may not be the most sensitive organisms to anthropogenic inputs. This paper describes preliminary results of toxicity tests with the mayfly, Isonychia bicolor (Ephemeroptera). Results suggested that Isonychia were moderately sensitive to NaCl after 96 h with an average LC₅₀ value of 3.10 g NaCl per liter. This value decreased after 7 days of exposure, resulting in a mean LC₅₀ value of 1.73 g NaCl per liter. When exposed to a coal-mine-processed effluent, Isonychia generated LC₅₀ values that ranged from 13% to 39% effluent. I. bicolor were more sensitive to the coal processing effluent than Ceriodaphnia dubia with conductivity lowest observable effects concentration (LOEC) values for mayfly survivorship that ranged from 1,508 to 4,101 μS/cm, while LOEC values for C. dubia reproduction ranged from 2,132 to 4,240 μS/cm.     

Toxicity of Surficial Sediments from Sydney Harbour and Vicinity, Australia

The toxicological responses of three species to 103 surficial saltwater sediment samples from Sydney Harbour, and coastal lakes and estuaries on the south-east coast of New South Wales, Australia, were tested in a battery of four to six laboratory toxicity tests. This is the first large-scale toxicological study of sediments in Australia, the objective of which is to assess the protective and predictive abilities of North American biological effects-based sediment quality guidelines, recently adopted in Australia. Amphipods were exposed to whole sediments in survival and reburial tests, sea urchin fertilisation and larval development tests were conducted on porewaters, and bacterial bio-luminescence (Microtox) tests were conducted on organic solvent extracts and porewaters. Local indigenous species were used for the amphipod and sea urchin tests (Corophium sp. and Heliocidaris tuberculata, respectively). A wide range of responses, from <25 to 100% of negative controls were observed in all tests. Mean control-adjusted responses ranged from 46 to 96% for all tests. The percentages of highly toxic samples ranged from 11 to 83% in the various tests. The order of test sensitivity was: amphipod survival < Microtox test of porewaters < amphipod reburial < sea urchin larval development < sea urchin fertilisation < Microtox test of solvent extracts. Concordance between toxicity tests in classifying samples as highly toxic or not, ranged from 47 to 79%, indicating some similarities between test results, but not complete equivalence. Combined toxicity test results showed that the incidence of highly toxic responses occurring in the majority of tests (75-100% of tests) was low (5% of samples), but a large percentage of samples had highly toxic results in at least one test (76% of samples). Toxicity was more pervasive in the Sydney region than in coastal lakes and estuaries south of Sydney. The current study demonstrated the utility of indigenous invertebrate species and the Microtox bacterium in a sediment toxicity test battery for Australian saltwater sediments.     

Seasonal availability and sensitivity of two field-collected mayflies for the development of a standardized toxicity test

Ecologically relevant toxicity tests may provide the best protection of sensitive aquatic fauna, but without established culturing or test methodology for such organisms, results may be unreliable and difficult to repeat. Further, field-collected organisms may not be feasible for routine testing purposes, as often required for permitted discharges. This study examined the feasibility of testing two field-collected mayflies, Isonychia bicolor and Maccaffertium spp., over a 1-year period. Seasonal comparisons of availability indicated I. bicolor and Maccaffertium spp. were most abundant during the winter months, resulting in 31 and 49 % of total organisms collected in 2009, while summer was the most difficult time to collect either species. Initial testing in January 2009 resulted in the highest no observable effect concentration (NOEC) values for survivorship (8 g NaCl for I. bicolor and 4 and 8 g NaCl/L for Maccaffertium spp.) when tested at 9°C. Subsequent tests conducted at 20–23°C resulted in 7-day NOEC values substantially lower (mean?=?1.44 and 1.59 g NaCl/L). Geometric means of exuviae indicated a dose-dependent response for I. bicolor exposed to NaCl, while no dose-dependent response was observed for Maccaffertium spp. with average number of molts varying from 4.93 in the 0.5 g NaCl/L concentration to 3.80 for control organisms followed by 2.24 (1 g NaCl/L). Averages again increased to 3.09 in the 2 g NaCl/L concentration, but declined in the highest concentrations (4–10 g NaCl/L). Based on the results of this feasibility study, field-collected mayflies appear to be too unpredictable in test responses, and therefore, such tests would be unreliable as stand-alone indicators of effluent toxicity.     

Application of a toxicity test battery integrated index for a first screening of the ecotoxicological threat posed by ports and harbors in the southern Adriatic Sea (Italy)

Ports and harbors may represent a threat for coastal ecosystems due to pollutant inputs, especially those derived from maritime activities. In this study, we report a first assessment of the ecotoxicological threat posed by six ports and harbors of opposite coastal regions, Apulia and Albania, in the southern Adriatic Sea (Italy). A bioassay battery consisting of four different species representing different trophic levels, algae Dunaliella tertiolecta, bacteria Vibrio fischeri, crustacean Artemia salina, and echinoids Paracentrotus lividus, has been used to assess sediment elutriates, pore waters, and sediment suspensions. Two different approaches of toxicity data integration, worst case and integrated index, have been used to determine the most appropriate procedure for the investigated sites. All sites with the worst case approach showed high toxicity levels. The chronic test with algae was the most sensitive identifying the highest effects in the battery. This effect can be attributable to contaminants derived from antifouling paints. The sediments, evaluated with V. fischeri test, often showed toxicity not found in the aqueous matrices of the same sites and that can be mainly linked to organic compounds. The test battery used in this study allowed us to perform a preliminary screening of the ecotoxicological risk of the studied area. In fact, the species utilized for toxicity tests responded differently to the investigated samples, showing different sensitivity. The test battery integrated index did not allow highlighting the differences among the sites and showed a general high ecotoxicological risk. A larger number of tests with higher sensitivity together with a tailored attribution of weights to endpoints and matrices will improve the final site evaluation.     

废水现状毒性试验(鱼类)方法的宏观质量控制因素分析

工业废水急性毒性污染的监测,以鱼类毒性试验的应用最为普及。与毒性达标控制的监测监管相比,毒性现状测定更有助于支持毒性污染预警监管的实施。通过对已知物质毒性试验结果的多次验证,证实由鱼类种类决定的方法检出限、实验中使用鱼类的种类形式、同种试验鱼的不同来源和依照"预实验"线性对现状进行的反推等4方面因素中除试验鱼的来源问题外,其余3因素已构成了对实验结果的决定性影响。3种宏观质量影响因素中,反推影响更难于实施质量控制。以现状表述为主要目标的毒性试验应尽可能回避稀释过程。     

The use of phytotoxicity tests (common duckweed,cabbage, and millet) for determining effluent toxicity

The objective of this study was to use higher plants for detecting effluent toxicity. Eight effluent samples were obtained from three industrial sources prior to their entry into a sewer system. The tests were the duckweed reproduction test, and root growth tests using cabbage and millet. The results of repeated phytotoxicity tests were reproducible. Of the three industrial sources, the effluent samples from a specialty chemical industry were the most toxic. For two samples from this source, the IC50 values (the concentrations which caused 50% inhibitory effect) for duckweed were less than 1.6% effluent concentration. The samples from an agricultural product utilization plant were the least toxic. For these samples, root growth tests failed to obtain IC50 values while the duckweed tests showed IC50 values of 91 and 43% effluent concentration. Among the three types of tests conducted, the duckweed reproduction test showed the greatest sensitivity to effluent toxicity, while root growth tests using cabbage and millet had mixed results. Duckweed is recommended as a part of a battery of tests for effluent toxicity.     

Validation of plant based bioassays for the toxicity testing of Indian waters

Plant-based bioassays have recently gained remarkable popularity among the toxicological/eco-toxicological assessment procedures. The reasons for their wide use are comparative simplicity, sensitivity, and cost-effectiveness as well as a good correlation with other toxicity tests. The present study describes the use of two plant bioassays, Allium cepa test and seed germination test in the evaluation of the toxicity/genotoxicity of industrial waste water and river water and standardization with the commonly occurring pollutants in Indian waters namely heavy metals and phenolics. Both tests were standardized to suit the Indian conditions, and the local varieties were used. Both bioassays responded significantly with the test range of heavy metals and phenolics. The toxicity of heavy metals was in the order of Cu > Ni > Cd in both the tests whereas 2,4-dinitrophenol was the most toxic among the phenolic compounds. Cabbage, millet, and cucumber, respectively, were found to be the most sensitive in the seed germination test for the test heavy metals and phenols. Significant amounts of chromosomal abnormalities including bridges, stickiness, and fragmentations were recorded with both the industrial waste water and the XAD concentrated river water samples by A. cepa test.     

Impacts of the fish farms on the water column nutrient concentrations and accumulation of heavy metals in the sediments in the eastern Aegean Sea (Turkey)

Pulp and paper mill effluents may cause harmful effects to the aquatic environment due to the combined influence of physical factors, toxic compounds, and nutrient enrichment. In the present study, the effectiveness of secondary treatment in reducing the toxicity of an elemental chlorine-free bleached-kraft pulp mill effluent was evaluated. To characterize the toxicity of the effluent, before and after the implementation of secondary treatment, a battery of tests with organisms bearing different functions at the ecosystem level was used, namely Vibrio fischeri (5-min luminescence), Pseudokirchneriella subcapitata (72-h growth), Lemna minor (7-day growth), Daphnia magna (21-day reproduction and 24-h postexposure feeding), Chironomus riparius (9-day growth), and Danio rerio (28-day growth). For the effluent sample collected before the implementation of secondary treatment, P. subcapitata was the most sensitive organism followed by V. fischeri and D. magna, and no toxic effects were observed toward the other organisms. For the effluent sample collected after the implementation of secondary treatment, the effluent caused no toxic effects on any of the tested species. The present results demonstrated not only that secondary treatment efficaciously reduced effluent toxicity toward the selected test organisms but also the usefulness of a battery of tests to characterize the toxicity of pulp mill effluents.     

Are single species toxicity tests alone adequate for estimating environmental hazard?

Most biologists agree that at each succeeding level of biological organization new properties appear that would not have been evident by even the most intense and careful examination of lower levels of organization. These levels might be crudely characterized as subcellular, cellular, organ, organism, population, multispecies, community, and ecosystem. The field of ecology developed because even the most meticulous study of single species could not accurately predict how several such species might interact competitively or in predator-prey interactions and the like. Moreover, interactions of biotic and abiotic materials at the level of organization called ecosystem are so complex that they could not be predicted from a detailed examination of isolated component parts. This preamble may seem platitudinous to most biologists who have heard this many times before. This makes it all the more remarkable that in the field of toxicity testing an assumption is made that responses at levels of biological organization above single species can be reliably predicted with single species toxicity tests. Unfortunately, this assumption is rarely explicitly stated and, therefore, often passes unchallenged. When the assumption is challenged, a response is that single species tests have been used for years and no adverse ecosystem or multispecies effects were noted. This could be because single species tests are overly protective when coupled with an enormous application factor or that such effects were simply not detected because there were no systematic, scientifically sound studies carried out to detect them. Probably both of these possibilities occur. However, the important factor is that no scientifically justifiable evidence exists to indicate the degree of reliability with which one may use single species tests to predict responses at higher levels of biological organization. One might speculate that the absence of such information is due to the paucity of reliable tests at higher levels of organization. This situation certainly exists but does not explain the lack of pressure to develop such tests. The most pressing need in the field of toxicity testing is not further perfection of single species tests, but rather the development of parallel tests at higher levels of organization. These need not be inordinately expensive, time consuming, or require any more skilled professionals than single species tests. Higher level tests merely require a different type of biological background. Theoretical ecologists have been notoriously reluctant to contribute to this effort, and, as a consequence, such tests must be developed by this and other organizations with similar interests.Paper presented at a Symposium held on 20–21 April 1982, in Edmonton, Alberta, Canada.     

Euro Chlor Risk Assessment for the Marine Environment Osparcom Region: North Sea - Chloroform

This risk assessment on chloroform was carried out specifically for the marine environment, according to the methodology laid down in the EU risk assessment Regulation (1488/94) and the Guidance Document of the EU New and Existing Substances Regulation (TGD, 1997). The study consists of the collection and evaluation of data on effects and environmental concentrations from analytical monitoring programs in large rivers and estuaries in the North Sea area. The risk is indicated by the ratio of the "predicted environmental concentrations" (PEC) and the "predicted no effect concentrations" (PNEC) for the marine aquatic environment. In total, 23 studies for fish, 17 studies for invertebrates and 10 studies for algae have been evaluated. Both acute and chronic toxicity studies have been taken into account and the appropriate assessment factors have been used to define a typical PNEC value of 72 µg/l. Due to limitations of the studies evaluated, a worst PNEC of 1 µg/l could also be used. Most of the available monitoring data apply to rivers and estuaries and were used to calculate PECs. The most recent data (1991-1995) support a typical PEC of 0.2 µg chloroform per litre of water and a worst case PEC of 5 to 11.5 µg chloroform per litre of water. The calculated PEC/PNEC ratios give a safety margin of 6 to 360 between the predicted no effect concentration and the exposure concentrations. A worst case ratio, however, points to a potential risk for sensitive species. Refinement of the assessment is necessary by looking for more data. Additional evaluation of environmental fate and bioaccumulation characteristics showed that no concern is expected for food chain accumulation.     

发光菌毒性试验在项目竣工环保验收监测中的应用

某敏感化工技改项目曾因超标排污引发跨地区厂群纠纷被责令停产整改,为考察该项目整改后废水处理设施的运行效果,保障水生生物安全,在该项目竣工环境保护验收监测时,采用发光细菌毒性试验检测废水的急性毒性.结果表明,达标排放的废水对生物的急性毒性为低毒,试验结果与常规验收监测结果有可比性,发光细菌毒性试验可用于必须公示的敏感项目竣工环境保护验收监测.     

Toxicity assessment of individual ingredients of synthetic-based drilling muds (SBMs)

Synthetic-based drilling muds (SBMs) offer excellent technical characteristics while providing improved environmental performance over other drilling muds. The low acute toxicity and high biodegradability of SBMs suggest their discharge at sea would cause minimal impacts on marine ecosystems, however, chronic toxicity testing has demonstrated adverse effects of SBMs on fish health. Sparse environmental monitoring data indicate effects of SBMs on bottom invertebrates. However, no environmental toxicity assessment has been performed on fish attracted to the cutting piles. SBM formulations are mostly composed of synthetic base oils, weighting agents, and drilling additives such as emulsifiers, fluid loss agents, wetting agents, and brine. The present study aimed to evaluate the impact of exposure to individual ingredients of SBMs on fish health. To do so, a suite of biomarkers [ethoxyresorufin-O-deethylase (EROD) activity, biliary metabolites, sorbitol dehydrogenase (SDH) activity, DNA damage, and heat shock protein] have been measured in pink snapper (Pagrus auratus) exposed for 21 days to individual ingredients of SBMs. The primary emulsifier (Emul S50) followed by the fluid loss agent (LSL 50) caused the strongest biochemical responses in fish. The synthetic base oil (Rheosyn) caused the least response in juvenile fish. The results suggest that the impact of Syndrill 80:20 on fish health might be reduced by replacement of the primary emulsifier Emul S50 with an alternative ingredient of less toxicity to aquatic biota. The research provides a basis for improving the environmental performance of SBMs by reducing the environmental risk of their discharge and providing environmental managers with information regarding the potential toxicity of individual ingredients.     

The importance of adult Hediste (Nereis) diversicolor in managing heavy metal pollution in shores and estuaries

The main aim of the toxicity test is to be able to measure toxicity in organisms, and therefore to assess whether a nationally or internationally preset standard is met. LC₅₀ values (median lethal concentration that kills 50% of the population at a given time) or LT₅₀ values (median lethal time in which 50% of the test species died) provide useful information for risk assessment. However, extrapolating laboratory data to actual field conditions is difficult because of changing environmental factors like temperature and salinity. LC₅₀ copper data for Hediste diversicolor treated multifactorially show the plasticity of the worm to environmental variables. Increasing temperature, 12 to 22°C, and increasing salinity, 7.6 to 30.5 reduced toxicity of copper to the worms without sediment. In the presence of sediment increasing temperature and increasing salinity increased toxicity of copper to the worms.     

Necessity of Toxicity Assessment in Turkish Industrial Discharges (Examples from Metal and Textile Industry Effluents)

Toxicity of some organic and inorganic chemicals to microorganisms is an important consideration in assessingtheir environmental impact against their economic benefits.Microorganisms play an important role in several environmentalprocesses, both natural and engineered. Some organic and inorganics at toxic levels have been detected in industrial discharges resulting in plant upsets and discharge permit violations. In addition to this, even though in some cases the effluent wastewater does not exceed the discharge limits,the results of toxicity tests show potential toxicity. Toxicityknowledge of effluents can benefit treatment plant operators inoptimising plant operation, setting pre-treatment standards, and protecting receiving water quality and in establishing sewer discharge permits to safeguard the plant. In the Turkish regulations only toxicity dilution factor (TDF) with fish is part of the toxicity monitoring program of permissible wastewater discharge. In various countries, laboratory studiesinvolving the use of different organisms and protocol for toxicity assessment was conducted involving a number of discharges.In this study, it was aimed to investigate the acute toxicity of textile and metal industry wastewaters by traditional and enrichment toxicity tests and emphasize the importance of toxicity tests in wastewater discharge regulations. The enrichment toxicity tests are novel applications and give anidea whether there is potential toxicity or growth limiting and stimulation conditions. Different organisms were used suchas bacteria (Floc and Coliform bacteria) algae (Chlorella sp.), fish (Lepistes sp.) and protozoan (Vorticella sp.) to represent four tropic levels. The textile industry results showed acute toxicity for at least one organism in 8 out of 23 effluent samples. Acute toxicity for at least two organisms in 7 out of 23 effluent sampling was observed for the metal industry. The toxicity test results were assessed with chemical analyses such as COD, BOD, color and heavy metals. It was observed that the toxicity of the effluents could not be explained by using physicochemical analyses in 5 cases for metal and 4 cases for the textile industries. The results clearly showed that the useof bioassay tests produce additional information about the toxicity potential of industrial discharges and effluents.     

Ecological risk assessment of perfluorooctanoic acid (PFOA) and perfluorooctanesulfonic acid (PFOS) in marine environment using Isochrysis galbana, Paracentrotus lividus, Siriella armata and Psetta maxima

Perfluorooctanoic acid (PFOA) and perfluorooctane sulfonic acid (PFOS) are anthropogenic substances classified as persistent bioaccumulative compounds and are found in various environmental compartments throughout the world, from industrialized regions to remote zones far from areas of production. In this study, we assessed the effects of PFOA and PFOS on early life stages of marine test species belonging to three different trophic levels: one microalga (Isochrysis galbana), a primary consumer (Paracentrotus lividus) and two secondary consumers (Siriella armata and Psetta maxima). Acute EC(50) values for PFOS were 0.11 mg L(-1) in P. maxima, 6.9 mg L(-1) in S. armata, 20 mg L(-1) in P. lividus and 37.5 mg L(-1) in I. galbana. In the case of PFOA, the toxicity was lower but the ranking was the same; 11.9 mg L(-1) in P. maxima, 15.5 mg L(-1) in S. armata, 110 mg L(-1) in P. lividus and 163.6 mg L(-1) in I. galbana. The Predicted No Effect Concentration (PNEC) for PFOS and PFOA in marine water derived from these acute toxicity values are 1.1 μg L(-1) for PFOS and 119 μg L(-1) for PFOA. This study established a baseline dataset of toxicity of PFOS and PFOA on saltwater organisms. The data obtained suggest that PFOA pose a minor risk to these organisms through direct exposure. In the perspective of risk assessment, early life stage (ELS) endpoints provide rapid, cost-effective and ecologically relevant information, and links should be sought between these short-term tests and effects of long-term exposures in more realistic scenarios.